The instant invention relates to a method for determining susceptibility to Escherichia coli urinary tract infections (UTI) and corresponding methods for diagnosing secretors and nonsecretors of histo-blood group antigens based on assaying urogenital cells and secretions. The instant invention also relates to methods and compositions for preventing and treating E. coli urinary tract infections. For example, disialosyl galactosyl-globoside is useful for diagnostic purposes and sialosyl galactosyl-globoside is useful for treatment and prevention of UTI.

Among the most common bacterial infections encountered in clinical practice are urogenital mucosal infections caused by E. coli, Chiamydlia trachomatis and N. gonorrheae which are diagnosed and treated at a cost to society in the billions of dollars. These infections are frequently recurrent and efforts at prevention have been largely fruitless, in part because of the apparent ineffectiveness of the mucosal immune system. Mucosal immunity is short-lived and in the long term, not protective. Despite much effort, attempts to develop vaccines against these mucosal urogenital pathogens have not succeeded and even natural immunity does not protect against recurrent episodes of infection with these agents. In addition, antigenic variability of the organisms has complicated vaccine development.
In particular, acute uncomplicated urinary tract infections occur in millions of young women each year. While most of these women experience only single or sporadic infections, approximately 20% suffer very frequent (xe2x89xa73/year) recurrences (Mabeck, C. E., Postgrad. Med J., 48:69-75, 1972). The apparent increased susceptibility to urinary tract infection in these patients cannot be explained by underlying functional or anatomic abnormalities of the urinary tract, but instead appears to arise from the interaction of infecting E. coli strains with these patients"" epithelial cells.
Thus, women prone to frequent recurrences demonstrate prolonged colonization of the vaginal mucosa with E. coli, the predominant causative species in these infections, (Stamey, T. A. et al., J. Urol, 113:214-217, 1975) and three-fold more E. coli adhere to vaginal, buccal and uroepithelial cells from women with recurrent urinary tract infection than to cells from control patients (Schaeffer, A. J. et al., N. Engl. J. Med., 304:1062-1066, 1981; Svanborg-Eden, C. et al., Infect. Immun., 26:837-840, 1979). Women with a history of recurrent urinary tract infections are also more likely to be nonsecretors of histo-blood group antigens than are women without a history of infections (relative risk=3 to 4 (Kinane, D. F. et al., Br. Med. J., 285:7-9, 1982; Sheinfeld, J. et al., N. Engl. J. Med., 320:773-777, 1989; Hooton, T. M. et al., Twenty-ninth Interscience Conference on Antimicrobial Agents and Chemotherapy, 8, 1989 (Program and Abstracts); Lomberg, H. et al., Infect. Immun., 51:919-926, 1986)), and E. coli adhere in greater numbers to uroepithelial cells from nonsecretors (Lomberg, H. et al., Infect. Immun., 51:919-926, 1986).
E. coli adhesins also play an important role in the pathogenesis of urinary tract infection (Stapleton, A. et al., J. Infect Dis., 163:773-779, 1989; Johnson, J. R., J. Infect. Dis., 156:225-229, 1987; Jacobson, S. H. et al., J. Infect. Dis., 152:426-427, 1985; Vaisenen, V. et al., Lancet, 2:1366-1369, 1981; Kallenius, G. et al., Lancet, 2:1369-1372, 1981; Vaisanen-Rhen, V. et al., Infect. Immun., 48:149-155, 1984; Westerlund, B. et al., J. Infect. Dis., 158:996-1002, 1988; Arthur, M. et al., Infect. Immun., 57:303-313, 1989; Sandberg, T., B. Kaijser, J. Clin. Microbiol, 26:1471-1476, 1988; O""Hanley, P. et al., N. Engl. J. Med., 313:414-420, 1985; Lidefelt, K.-J. et al., Acta Paediatr. Scand., 76:775-780, 1987; Lund, B. et al., Mol. Microbiol., 2:255-263, 1988), particularly the genetically related P and F adhesins.
P adhesins are present in 50% to 65% of E. coli strains causing cystitis (Stapleton, A. et al., J. Infect Dis., 163:773-779, 1989; Arthur, M. et al., Infect. Immun., 57:303-313, 1989; Sandberg, T., B. Kaijser, J. Clin. Microbiolp, 26:1471-1476, 1988; O""Hanley, P. et al., N. Engl. J. Med., 313:414-420, 1985; Lidefelt, K.-J. et al., Acta Paediatr. Scand., 76:775-780, 1987; Lund, B. et al., Mol. Microbiol., 2:255-263, 1988) and 75% to 90% of isolates from pyelonephritis (Johnson, J. R., J. Infect. Dis., 156:225-229, 1987; Jacobson, S. H. et al., J. Infect. Dis., 152:426-427, 1985; Vaisenen, V. et al., Lancet, 2:1366-1369, 1981; Kallenius, G. et al., Lancet, 2:1369-1372, 1981; Vaisanen-Rhen, V. et al., Infect. Immun., 48:149-155, 1984; Westerlund, B. et al., J. Infect. Dis., 158:996-1002, 1988; Arthur, M. et al., Infect. Immun., 57:303-313, 1989; Sandberg, T., B. Kaijser, J. Clin. Microbiol, 26:1471-1476, 1988). The minimal receptor for the P adhesin is the galactose xcex11-4 galactose moiety, present in the globoseries glycolipids and the P1 blood group antigen (Kallenius, G. et al., FEMS Microbiol Lett., 7:297-302, 1980; Leffler, H. et al., FEMS Microbiol. Lett., 8:127-134, 1980; Svanborg-Eden, C. et al., Scand. J Infect Dis. Suppl., 24:144-147, 1980; Kallenius, G. et al., Infection, 8 (Suppl. 3):S288-S293, 1981; Kallenius, G. et al., Lancet, 2:604-606, 1981; Leffler, H. et al., Infect Immun., 34:920-929, 1981; Kallenius, G. et al., Scand. J. Infect. Dis. Suppl., 33:52-60, 1982; Svenson, S. B. et al., Infection, 11:73/61-79/67, 1983; Bock, K. et al., J. Biol Chem., 260:8545-8551, 1985).
F adhesins are expressed in 30% to 65% of urinary tract infection isolates (Stapleton, A. et al., J. Infect Dis., 163:773-779, 1989; Arthur, M. et al., Infect. Immun., 57:303-313, 1989). The minimal binding moiety for the F adhesin is less well defined and probably more complex; proposed receptors include the Forssman and para-Forssman antigens, globoside, galactosyl globoside, globo A and globo H, and stage-specific embryonic antigen-4 (SSEA-4) (Lund, B. et al., Mol. Microbiol., 2:255-263, 1988; Lindstedt, R. et al., Infect. Immun., 57:3389-3394, 1989; Karr, J. F. et al., Infect Immun., 58:4055-4062, 1990; Stromberg, N. et al., EMBO J., 9:2001-2010, 1990). The globoseries glycolipids have recently been shown to be modified by the histo-blood group status, and ABH active globoseries GSLs are found in renal epithelium and in voided uroepithelial cells as well as on erythrocytes (Leffler, H. et al., FEMS Microbiol. Lett., 8:127-134, 1980; Bock, K. et al., J. Biol Chem., 260:8545-8551, 1985; Breimer, M. E. et al., FEBS Lett, 179:165-172, 1985; Breimer, M. E. et al., J. Biochem., 98:1169-1180, 1985; Lindstedt, R. et al., Infect. Immun., 59:1086-1092, 1991; Clausen, H. et al., Vox Sang., 56:1-20, 1989; Kannagi, R. et al., FEBS Lett., 175:397-401, 1984; Clausen, H. et al., Biochem. Biophys. Res. Commun., 124:523-529, 1984).
Several lines of evidence have suggested that the increased susceptibility to recurrent urinary tract infection observed in some otherwise healthy women may be explained by genetic factors influencing the density and/or specificity of bacterial receptors available to mediate colonization of their uroepithelial cells (Schaeffer, A. J. et al., N. Engl. J. Med., 304:1062-1066, 1981; Svanborg-Eden, C. et al., Infect. Immun., 26:837-840, 1979; Kinane, D. F. et al., Br. Med. J., 285:7-9, 1982; Sheinfeld, J. et al., N. Engl. J. Med., 320:773-777, 1989; Hooton, T. M. et al., Twenty-ninth Interscience Conference on Antimicrobial Agents and Chemotherapy, 8, 1989 (Program and Abstracts); Lomberg, H. et al., Infect. Immun., 51:919-926, 1986). Although it is known that nonsecretors of histo-blood group antigens are overrepresented among women with a history of recurrent urinary tract infections (Kinane, D. F. et al., Br. Med. J., 285:7-9, 1982; Sheinfeld, J. et al., N. Engl. J. Med., 320:773-777, 1989; Hooton, T. M. et al., Twenty-ninth Interscience Conference on Antimicrobial Agents and Chemotherapy, 8, 1989 (Program and Abstracts)) and that uroepithelial cells from nonsecretors show enhanced E. coli adherence compared with cells from ,sedcretors (Lomberg, H. et al.,. Infect. Immun., 51:919-926, 1986), the biochemical basis for these observations has not been clarified.
Accordingly, an object of the present invention is to determine the biochemical basis for the overrepresentation of nonsecretors among women with a history of recurrent E. coli urinary tract infections. Another object of the present invention is to provide a method for determination of susceptibility to E. coli urinary tract infections as well as methods for diagnosing secretors and nonsecretors. Another object of the instant invention is to provide methods and compositions for preventing and treating UTI. A preferred composition comprises SGG or derivatives thereof, particularly stabilized forms.
Accordingly, the present invention relates to a method for determining susceptibility to E. coli urinary tract infection comprising assaying a sample of epithelial cells for the presence or absence of at least one of Lewisa (Lea), sialosyl galactosyl-globoside (sialosyl gal-globoside or SGG), disialosyl galactosyl-globoside (disialosyl gal-globoside or DSGG) and an extended globo structure carrying the same terminal epitopes as Lea, SGG or DSGG or assaying a sample of vaginal secretions for the presence or absence of at least one of SGG or DSGG, and detecting the presence or absence of the at least one of Lea, SGG, DSGG and the extended globo structure.
A preferred embodiment relates to the use of SGG, or stabilized forms thereof, to intervene in the attachment of E. coli to urogenital epithelium. A suitable stabilized form of SGG is where the sialic acid residue is replaced by 2-keto-3-deoxy-D-glycero-D-galacto-nonulonic acid, otherwise known as KDN. KDN essentially is deaminated neuraminic acid. KDN can be incorporated readily into a synthetic scheme for making SGG using KDN transferase to add KDN to the terminal Gal residue.
The present invention also relates to a medicament comprising a biologically effective amount of at least one E. coli bacterial receptor analogue, and a pharmaceutically acceptable diluent, carrier or excipient as well as a method for preventing E. coli urinary tract infection comprising administering, to a host, a biologically effective amount of at least one E. coli bacterial receptor or bacterial receptor analogue.